Purification and properties of RNA polymerases from mother cells and forespores of sporulating cells of Bacillus subtilis.
Bacillus subtilis sporulating cells at stage III were fractionated into mother cell and forespore fractions by means of a lysozyme-detergent method. Three forms of DNA-dependent RNA polymerase enzymes, termed M sigma, F sigma, and F delta, in addition to core enzyme (alpha 2, beta', and beta) have been purified from the cell fractions. Enzymes M sigma and F sigma are present in the mother cell and forespore, respectively, and contain sigma factor of 55,000 daltons in addition to the core subunits. On the other hand, enzyme F delta is present specifically in the forespore and contains delta 1 factor of 28,000 daltons instead of the sigma factor. The amount of RNA polymerase in the forespore is about twice that in the mother cell. The enzymes M sigma and F sigma also differed in their elution profiled from DEAE-cellulose columns and in their heat stabilities indicating that the two sigma-containing holoenzyme forms may be different in their structural properties. The enzyme F delta transcribed B. subtilis DNA about 1.6 times more actively than enzyme F sigma, and the enzymes M sigma and F sigma transcribed the DNA about 2.2 times more actively than did core enzyme.
D001412:Bacillus subtilis / Q000201:enzymology* / Q000254:growth & development; D004269:DNA, Bacterial / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification* / Q000378:metabolism; D004355:Drug Stability; D006358:Hot Temperature; D046911:Macromolecular Substances; D011067:Poly dA-dT / Q000378:metabolism; D013171:Spores, Bacterial / Q000201:enzymology; D013698:Templates, Genetic; D014158:Transcription, Genetic
D004269:DNA, Bacterial; D046911:Macromolecular Substances; D011067:Poly dA-dT; D012321:DNA-Directed RNA Polymerases